The experimental Immunology Core Facility will provide expertise in state- of-the-art immunological procedures for used by basic and clinical CFAR investigators. Essential assays for immunology research will be provided on a fee-for-service basis. The fee structures will include a partial subsidy from the core grant, which will enable highly qualified technical personnel to be available to participate in numerous research projects. Additional objectives include training of personnel from other laboratories, and serving as a resource to provide immunological reagents to the CFAR community. The long term objectives are to facilitate research and promote collaboration among CFAR investigators. Individual components of this Core Facility include: (a) Molecular Immunology. The laboratory has developed unique expertise and reagents for analysis of cytokines, chemokines, chemokine receptors, and growth factors in non-human primates as well as human cells and tissues. (B) Flow cytometry. New flow cytometry methods have recently been introduced for detection of specific cell- mediated immune responses to HIV, including labeling of antigen-specific cells with soluble MHC-tetramers and detections of intracellular cytokines produced by specifically stimulated cells. The goal of the Emory Center for AIDS Research Flow Cytometry Facility will be to provide these new techniques, as well as traditional immunophenotyping analysis, to clinical and basic studies investigating HIV-specific immune responses in infected humans, and to apply them in primate models used for pathogenesis studies and vaccine development. (C) Evaluation of Humoral Immune Responses. This laboratory will provide standardized assays for analysis include antibody- capture ELISAs for the detection of HIV/SIV anti-gag and anti-env antibodies, CEM x 174 co-culture to detect serum neutralizing antibody responses against various HIV and SIV strains, and Western blots to examine the breadth of serologic response against HIV or SIV. (D) Confocal Microscopy and Immunohistochemistry. The confocal microscopy and immunohistochemistry facility will enable CFAR members to obtain high resolution, digital confocal images of cells. Examples of applications include localization and co-localization of viral antigens in infected cells, identification of subcellular compartments, containing viral proteins, assays of effects of localization of cellular proteins after different stimuli (e.g. capping of cell surface proteins when cells are activated), and localization of viral nucleic acids.